Analysis gene diversity of phenol hydroxylase and catechol 1,2-dioxygenase from fecal microbiome of Nycticebus pygmaeus

Abstract:

[Objective] The gene diversity of phenol hydroxylase and catechol 1,2-dioxygenase were investigated from fecal microbiome of Nycticebus pygmaeus. [Methods] Degenerate primers were used to amplify phenol hydroxylase and catechol 1,2-dioxygenase gene fragments from metagenomic DNA. Phenol hydroxylase and catechol 1,2-dioxygenase gene clone libraries were constructed, and some of clones were sequenced separately. [Results] The BLAST analysis of phenol hydroxylase and catechol 1,2-dioxygenase sequences showed 92%?100% and 87%?100% identities to the known phenol hydroxylase and catechol 1,2-dioxygenase sequences in GenBank. Phylogenetic analysis showed that phenol hydroxylase sequences in gene clone libraries had high similarity with phenol hydroxylase sequences from Neisseria, Burkholderia, Alcaligenes, Acinetobacter. And catechol 1,2-dioxygenase sequences in gene clone libraries had high similarity with catechol 1,2-dioxygenase sequences from Acinetobacter. Sequence alignment showed two DEXRH motifs of LmPH sequences were detected in phenol hydroxylase sequences, and the conserved cysteine was detected in catechol 1,2-dioxygenase sequences which was inhibited by Ag+ and Hg2+. [Conclusion] The phenol hydroxylase from fecal microbiome of Nycticebus pygmaeus was multicomponent phenol hydroxylase, and catechol that middle production of phenol degradation can be cleaved by catechol 1,2-dioxygenase through ortho-pathway.